Mechanism of lncRNA DUXAP8 in promoting proliferation of bladder cancer cells by regulating PTEN.

OBJECTIVE: To evaluate the lncRNA DUXAP8 expression in bladder cancer and its mechanism. PATIENTS AND METHODS: Clinical specimens were analyzed. The expression of lncRNA in bladder cancer and adjacent tissues was detected using qRT-PCR. The χ2-test analysis was used to analyze the relationship between lncRNA DUXAP8 expression and clinicopathological information in patients with bladder cancer. The tumor cell activity and cell proliferation were measured by cell counting kit-8 (CCK8) and colony formation assay. We utilized polymerase chain reaction (PCR) to access PTEN expression in bladder cancer and adjacent tissues. Pearson correlation analysis was utilized for evaluating the relationship between PTEN and lncRNA DUXAP8. Western blot was used for detecting protein expression. RESULTS: LncRNA DUXAP8 expression was higher in bladder cancer tissues; it was in a positive correlation with the TNM stage and tumor size, but negatively correlated with the total survival time. Knockdown of DUXAP8 decreased cell viability and cellular proliferation. Lower expression of PTEN gene was found in bladder cancer compared with that in adjacent tissues. Pearson correlation analysis showed that PTEN was negatively correlated with DUXAP8; knockdown of DUAP8 increased the expression of PTEN. Overexpressing DUAP8 increased protein level of PTEN, but decreased cell viability. CONCLUSIONS: Our results pointed out that lncRNA DUXAP8 was overexpressed in bladder cancer tissues, which can promote the progression of bladder cancer through inhibiting PTEN.

Delineation of an enhancerlike positive regulatory element in the interleukin-2 receptor alpha-chain gene.

We have delineated a positive regulatory element in the interleukin-2 receptor alpha-chain gene (IL-2R alpha) between positions -299 and -243 that can potently activate a heterologous (herpesvirus thymidine kinase [tk]) promoter in phorbol myristate acetate (PMA)-induced Jurkat T cells and is functional when cloned in either orientation. This enhancerlike element contains a site (-268/-257) that can bind NF-kappa B; however, unlike the immunoglobulin kappa gene kappa B enhancer element, the IL-2R alpha kappa B-like site alone can only weakly activate a heterologous promoter. Adjacent 5' and 3' sequences also weakly activate the tk-CAT vector, but constructs combining the IL-2R alpha kappa B-like site plus adjacent 5' and 3' sequences potently activate gene expression. This combination of regions is essential for potent PMA-induced transcription from the tk promoter. Experiments using constructs in which IL-2R alpha upstream sequences are sequentially deleted suggested that there is a region 5' of position -299 which can suppress IL-2R alpha promoter and/or enhancer activity. Thus, it is possible that both positive and negative elements may be important in the regulation of IL-2R alpha gene transcription.

In-vivo effects of amrinone and milrinone on left ventricular dP/dt, intraventricular pressure and heart rate in pentobarbital-anesthetized guinea pigs.

The cardiovascular properties of amrinone and milrinone were investigated in pentobarbital-anesthetized guinea pigs (in vivo). A 19g needle, attached to a pressure transducer, was inserted through the chest wall into the left ventricle. Continuous monitoring of intraventricular pressure and heart rate was recorded. The first derivative of this pressure pulse was dP/dt, an index of inotropic state. At a dose of 500 ug/Kg, amrinone given intravenously caused an increase in the rate of rise in left ventricular pressure (LV dP/dt) by 28.7 +/- 7.8% (from 928 +/- 34 to 119 +/- 73 mm Hg/sec., p less than 0.05). At 50 ug/Kg, milrinone (i.v.) caused an increase in the left ventricular peak positive dP/dt by 15.3 +/- 3.4 (from 942 +/- 24 to 1086 +/- 32 mm Hg/sec., P less than 0.001). Both amrinone and milrinone, in addition to vasodilating effect, exerted a concentration-related manner with these positive inotropic actions.

Digital photography for mapping Mohs micrographic surgery sections.

BACKGROUND: Instant photographic print images have been used to diagram tissue sections excised for Mohs micrographic surgery (MMS). This approach is limited by the size of the print image, the potential difficulty of writing on a glossy photo print, and the cost of film. OBJECTIVE: We describe the use of digital photographic images as templates for making maps of tissue excised for MMS. METHODS: Digital photographic images of patients undergoing MMS are downloaded to a computer and printed onto plain paper. A map of the tissue excised for MMS is drawn directly onto the digital print. RESULTS: Several methods of creating MMS maps using digital photographic print images are described. CONCLUSION: Advantages of using digital photographs in MMS include speed of producing images, low cost of materials, greater accuracy of depicting the MMS excision defect, and ease of correlating the MMS map to the patient for subsequent stages of excision.

Regulation of adenosine kinase by adenosine analogs.

The regulation of adenosine phosphorylation by adenosine analogs was studied using highly purified human placental adenosine kinase [ATP: adenosine 5'-phosphotransferase (EC 2.7.1.20)]. Our observations lead us to classify the analogs into three groups as follows: type I, 5'-N-ethylcarboxamidoadenosine and 5'-methylthioadenosine; type II, N6-cyclohexyladenosine, N6-L-phenylisopropyladenosine, and 2-chloroadenosine; and type III, 6-methylmercaptopurine riboside. Type I compounds are inhibitors of adenosine kinase at 0.5 microM adenosine with IC50 values of 25 microM for 5'-N-ethylcarboxamidoadenosine and 250 microM for 5'-methylthioadenosine. These compounds stimulate adenosine kinase at 5.0 microM adenosine up to a maximum of 30 to 50% above basal velocity. They are not substrates for adenosine kinase. Type II compounds are inhibitors of adenosine kinase at 0.5 microM adenosine with an IC50 of 220 microM for N6-cyclohexyladenosine and 200 microM for N6-L-phenylisopropyladenosine. These analogs also stimulate adenosine kinase at 5.0 microM adenosine. 2-Chloroadenosine, N6-cyclohexyladenosine, and N6-L-phenylisopropyladenosine are phosphorylated by adenosine kinase with apparent Km values of 1,330, and 205 microM, respectively. 6-Methylmercaptopurine riboside (type III) inhibited enzyme activity with an IC50 of 10 microM at 0.5 microM adenosine and 215 microM at 5 microM adenosine and is a substrate for adenosine kinase. These data are consistent with the following: (a) 2-chloroadenosine, N6-cyclohexyladenosine, and N6-L-phenylisopropyladenosine may not be good adenosine receptor agonists in vivo because they are phosphorylated into active derivatives by adenosine kinase; (b) 5'-N-ethylcarboxamidoadenosine and 5'-methylthioadenosine are superior candidates for adenosine receptor agonists in vivo because they are not phosphorylated; (c) 5'-N-ethylcarboxamidoadenosine, 5'-cyclohexyladenosine, N6-L-phenylisopropyladenosine, and 2-chloroadenosine may interact with adenosine kinase at two sites on the enzyme, a catalytic site and a regulatory site; and (d) 6-methylmercaptopurine riboside may interact with the enzyme at the catalytic site only.

The same target sequences are differentially important for activation of the interleukin 2 receptor alpha-chain gene in two distinct T-cell lines.

High-affinity receptors for interleukin 2 (IL-2) are expressed on T cells following activation. These receptors are composed of both alpha and beta chains. Expression of alpha chains and, therefore, expression of high-affinity receptors are critically regulated at the level of transcription initiation. We have further dissected the regulatory elements involved in controlling transcription of the IL-2 receptor alpha-chain (IL-2R alpha) gene. The IL-2R alpha promoter contains a kappa B site and binding sites for additional nuclear factors within a 50-base-pair region (positions -290 to -240 relative to the major transcription start site). These include one upstream of the kappa B site and one similar to the c-fos serum response element (SRE), which is downstream of the kappa B site. Mutation of the kappa B site decreases IL-2R alpha promoter activity in MT-2 cells (a T-cell line that has been transformed with human T-cell lymphotropic virus type I), but not in Jurkat cells (a T-cell leukemia line) that have been activated by phorbol 12-myristate 13-acetate (PMA). In contrast, mutation of a region upstream of the kappa B site decreases activity in PMA-induced Jurkat cells but increases activity in MT-2 cells. Mutation of the SRE-like site decreases activity in both cell types but the effect in PMA-induced Jurkat is more pronounced. Thus, these distinct cis-acting elements play different physiological roles in IL-2R alpha gene activation in MT-2 cells and PMA-induced Jurkat T cells. These studies provide direct evidence for a functionally significant SRE-like sequence in a gene other than c-fos and the actin genes and identify other elements that are critical for IL-2R alpha gene expression.

Aminoglycoside-resistance mechanisms for cystic fibrosis Pseudomonas aeruginosa isolates are unchanged by long-term, intermittent, inhaled tobramycin treatment.

Aminoglycoside-resistance mechanisms were characterized in Pseudomonas aeruginosa isolates from cystic fibrosis (CF) patients during a recent clinical trial of inhaled tobramycin. Impermeability, in which bacteria have reduced susceptibility to all aminoglycosides, was the predominant mode of resistance in isolates obtained both before and after 6 months of cyclic treatment with tobramycin or placebo administered by aerosol. Enzymatic resistance mechanisms were found in fewer than 10% of resistant isolates. P. aeruginosa from individual patients could be grouped on the basis of genetic relatedness. When enzymatic resistance was involved, all isolates in a group had elevated tobramycin MICs. When impermeability occurred, MICs of a genotypic group varied from susceptible to resistant. These findings suggest that impermeability resistance occurs in only a fraction of the P. aeruginosa population in lungs of persons with CF and that this form of resistance arises by a process involving multiple small changes in MIC.